Generation or large numbers of immature and mature dendritic cells from rat bone marrow cultures

We have defined conditions for generating large numbers of dendritic cells (DC) in marrow cultures from 10 – 12‐week‐old ACI or WF rats. The combination of granulocyte‐macrophage colony‐stimulating factor (GM‐CSF) and TNF‐α, known to induce DC from human CD34+ progenitors, was not effective with rat. In contrast, GM‐CSF plus IL‐4 generated DC in high yield, corresponding to 30 – 40 % of the initial number of plated marrow cells. The DC proliferated in distinctive aggregates, in which most cells had an immature phenotype marked by undetectable surface B7 and high levels of MHC class II products within intracellular lysosomes. When dislodged and dispersed, the aggregates gave rise to mature stellate DC with abundant surface MHC class II and B7, sparse MHC class II– lysosomes, and strong T cell‐stimulating capacity. Therefore, rat marrow progenitors can generate large numbers of immature DC, with abundant intracellular MHC class II compartments, and potent, stimulatory, mature DC.

[1]  R. Steinman,et al.  Developmental regulation of MHC class II transport in mouse dendritic cells , 1997, Nature.

[2]  P. Cresswell,et al.  HLA-DM is localized to conventional and unconventional MHC class II-containing endocytic compartments. , 1996, Immunity.

[3]  C Danieli,et al.  Dendritic cells use macropinocytosis and the mannose receptor to concentrate macromolecules in the major histocompatibility complex class II compartment: downregulation by cytokines and bacterial products , 1995, The Journal of experimental medicine.

[4]  M. Moore,et al.  Expansion of immunostimulatory dendritic cells among the myeloid progeny of human CD34+ bone marrow precursors cultured with c-kit ligand, granulocyte-macrophage colony-stimulating factor, and TNF-alpha. , 1995, Journal of immunology.

[5]  T. Starzl,et al.  A new protocol for the propagation of dendritic cells from rat bone marrow using recombinant GM-CSF, and their quantification using the mAb OX-62. , 1995, Journal of immunological methods.

[6]  R. Steinman,et al.  Proliferating dendritic cell progenitors in human blood , 1994, The Journal of experimental medicine.

[7]  F. Sallusto,et al.  Efficient presentation of soluble antigen by cultured human dendritic cells is maintained by granulocyte/macrophage colony-stimulating factor plus interleukin 4 and downregulated by tumor necrosis factor alpha , 1994, The Journal of experimental medicine.

[8]  R. Steinman,et al.  Generation of large numbers of dendritic cells from mouse bone marrow cultures supplemented with granulocyte/macrophage colony-stimulating factor , 1992, The Journal of experimental medicine.

[9]  J. Banchereau,et al.  GM-CSF and TNF-α cooperate in the generation of dendritic Langerhans cells , 1992, Nature.

[10]  J. Tikerpae,et al.  Interactions of tumor necrosis factor with granulocyte-macrophage colony-stimulating factor and other cytokines in the regulation of dendritic cell growth in vitro from early bipotent CD34+ progenitors in human bone marrow. , 1992, Journal of immunology.

[11]  R. Steinman,et al.  Identification of proliferating dendritic cell precursors in mouse blood , 1992, The Journal of experimental medicine.

[12]  G. Macpherson Properties of lymph-borne (veiled) dendritic cells in culture. I. Modulation of phenotype, survival and function: partial dependence on GM-CSF. , 1989, Immunology.

[13]  R. Willemze,et al.  Inhibition of human macrophage colony formation by interleukin 4 , 1989, The Journal of experimental medicine.

[14]  R. Steinman,et al.  Presentation of exogenous protein antigens by dendritic cells to T cell clones. Intact protein is presented best by immature, epidermal Langerhans cells , 1989, The Journal of experimental medicine.

[15]  M. Berkowitz,et al.  Differentiation of dendritic cells in cultures of rat bone marrow cells , 1986, The Journal of experimental medicine.

[16]  A. Helenius,et al.  Glycoproteins of the lysosomal membrane , 1985, The Journal of cell biology.

[17]  Jon,et al.  ACCESSORY AND STIMULATING PROPERTIES DENDRITIC CELLS AND MACROPHAGES ISOLATED FROM VARIOUS RAT TISSUES* OF BY WOLFGANG , 1982 .

[18]  M. Moore The clinical use of colony stimulating factors. , 1991, Annual review of immunology.